本文關(guān)鍵詞： 黃瓜 赤霉素受體 果實(shí)心室 瓠果　出處：《中國(guó)農(nóng)業(yè)大學(xué)》2017年博士論文　論文類型：學(xué)位論文

【摘要】：植物激素赤霉素(GA)是高等植物體內(nèi)的一類四環(huán)二萜羧酸化合物,在植物各個(gè)生長(zhǎng)發(fā)育階段起著至關(guān)重要的作用。前人已在赤霉素促進(jìn)種子發(fā)芽,調(diào)控植物營(yíng)養(yǎng)生長(zhǎng)和生殖生長(zhǎng)等方面開展了較多研究,但有關(guān)其調(diào)控植物果實(shí)發(fā)育方面的分子機(jī)制尚不明確。本文以黃瓜為試材,通過時(shí)空表達(dá)、功能驗(yàn)證和表達(dá)譜分析,研究了 GA受體蛋白CsGID1a對(duì)黃瓜果實(shí)心室形成的調(diào)控機(jī)制,旨在豐富黃瓜果實(shí)發(fā)育的理論基礎(chǔ),取得的主要結(jié)果如下:(1)黃瓜CsGID1a和CsGIID1b屬于高等植物赤霉素受體基因GID1家族的同源基因,均含有受體基因所特有的HGG和GXSXG兩個(gè)保守域,在黃瓜的莖尖和根尖處高度表達(dá),外源GA3處理可導(dǎo)致反饋調(diào)節(jié),使CsGID1a和CsGID1b的表達(dá)量下調(diào),表明CsGID1a和CsGID1b均具備赤霉素受體基因的典型特征。(2)進(jìn)化樹分析表明CsGID1a處于雙子葉植物GID1主效基因ac族,時(shí)空表達(dá)分析顯示CsGID1a在根尖、莖尖、雌花芽及果實(shí)中的表達(dá)顯著高于CsGID1b,干擾CsGID1b表達(dá)并不會(huì)影響植株生長(zhǎng),表明CsGID1a可能是主效基因。(3)通過原位雜交和GUS染色在花和果實(shí)中對(duì)CsGID1a的表達(dá)進(jìn)行更詳細(xì)分析表明,CsGID1a在黃瓜雄蕊、雌蕊及果實(shí)的心室壁上特異表達(dá)。通過黃瓜遺傳轉(zhuǎn)化獲得的CsGID1a干擾轉(zhuǎn)基因植株,與對(duì)照相比,干擾植株明顯矮化,果實(shí)心室發(fā)育不完全。表達(dá)譜分析表明在干擾轉(zhuǎn)基因植株中,生長(zhǎng)素合成與運(yùn)輸基因和細(xì)胞周期相關(guān)基因的表達(dá)量都顯著低于對(duì)照組;同時(shí)激素測(cè)定與細(xì)胞切片觀察的結(jié)果表明,CsGID1a干擾轉(zhuǎn)基因植株果實(shí)中生長(zhǎng)素的分布和果實(shí)細(xì)胞分化受到影響。(4)在擬南芥中的功能驗(yàn)證發(fā)現(xiàn),異源表達(dá)CsGID1a可部分恢復(fù)雙突變體gid1agid1c的植株高度以及植物育性表型,同時(shí)也對(duì)果實(shí)心室的發(fā)育造成了影響,部分?jǐn)M南芥轉(zhuǎn)基因株系的果實(shí)還形成了類似黃瓜果實(shí)的三心室。
[Abstract]:The plant hormone gibberellin (GA) is a kind of tetracyclic diterpenoid carboxylic acid compounds in higher plants, which plays an important role in the growth and development of plants. More studies have been carried out on the regulation of plant growth and reproductive growth, but the molecular mechanism of regulating plant fruit development is not clear. In this paper, cucumber was used as the material and expressed in time and space. The regulation mechanism of GA receptor protein CsGID1a on the ventricular formation of cucumber fruit was studied in order to enrich the theoretical basis of cucumber fruit development. The main results are as follows: (1) Cucumber CsGID1a and CsGIID1b belong to the GID1 family of gibberellin receptor genes in higher plants. There were two conserved domains, HGG and GXSXG, which were specific to the receptor gene. They were highly expressed at the stem tip and root tip of cucumber. Exogenous GA3 treatment could lead to feedback regulation. The expression of CsGID1a and CsGID1b was down-regulated. The results showed that both CsGID1a and CsGID1b had the typical character of gibberellin receptor gene. The phylogenetic tree analysis showed that CsGID1a was in the ac group of GID1 major genes in dicotyledonous plants. Spatio-temporal expression analysis showed that the expression of CsGID1a in root tip, stem tip, female flower bud and fruit was significantly higher than that in CsGID1b. Interfering with CsGID1b expression did not affect plant growth. The expression of CsGID1a in flower and fruit was analyzed in more detail by in situ hybridization and GUS staining. CsGID1a was specifically expressed in the ventricular wall of stamen, pistil and fruit of cucumber. CsGID1a interference transgenic plants obtained from cucumber genetic transformation were significantly dwarfed compared with the control. The expression profile analysis showed that the expression of auxin synthesis and transporter genes and cell cycle related genes were significantly lower in the transgenic plants than in the control group. At the same time, the results of hormone determination and cell section observation showed that CsGID1a interfered with the distribution of auxin and the differentiation of fruit cells in transgenic plants, and the function of CsGID1a was verified in Arabidopsis thaliana. The heterologous expression of CsGID1a could partially restore the plant height and fertility phenotype of the double mutant gid1agid1c, and also affect the development of fruit ventricle. The fruits of some Arabidopsis transgenic lines also formed three ventricles similar to cucumber fruits.
【學(xué)位授予單位】：中國(guó)農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：S642.2

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本文編號(hào)：1472574